Comparison of expression systems for the production of human interferon-a2b.

The production of human interferon alpha2b (IFN-α2b) in two expression systems, tobacco (Nicotiana tabaccum) and Escherichia coli, was compared in various aspects such as safety, yield, quality of product and productivity. In the E. coli system, IFN-α2b was expressed under a pelB signal sequence and...

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Main Authors: Memari, Hamid Rajabi, Ramanan, Ramakrishnan Nagasundara, Ariff, Arbakariya
Format: Article
Language:English
English
Published: SP Versita 2010
Online Access:http://psasir.upm.edu.my/id/eprint/13689/
http://psasir.upm.edu.my/id/eprint/13689/1/Comparison%20of%20expression%20systems%20for%20the%20production%20of%20human%20interferon.pdf
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author Memari, Hamid Rajabi
Ramanan, Ramakrishnan Nagasundara
Ariff, Arbakariya
author_facet Memari, Hamid Rajabi
Ramanan, Ramakrishnan Nagasundara
Ariff, Arbakariya
author_sort Memari, Hamid Rajabi
building UPM Institutional Repository
collection Online Access
description The production of human interferon alpha2b (IFN-α2b) in two expression systems, tobacco (Nicotiana tabaccum) and Escherichia coli, was compared in various aspects such as safety, yield, quality of product and productivity. In the E. coli system, IFN-α2b was expressed under a pelB signal sequence and a T7lac promoter in a pET 26b(+) vector. The same gene was also cloned in expression plant vector (pCAMBIA1304) between cauliflower mosaic virus promoter (CaMV35S) and poly A termination region (Nos) and expressed in transgenic tobacco plants. The expression of protein in both systems was confirmed by western immunoblotting and the quantity of the protein was determined by immunoassay. The amount of periplasmic expression in E. coli was 60 µg/L of culture, while the amount of nuclear expression in the plant was 4.46 µg/kg of fresh leaves. The result of this study demonstrated that IFN-α2b was successfully expressed in periplasm of bacterial and plant systems. The limitations on the production of IFN-α2b by both systems are addressed and discussed to form the basis for the selection of the appropriate expression platform.
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spelling upm-136892015-10-21T00:09:25Z http://psasir.upm.edu.my/id/eprint/13689/ Comparison of expression systems for the production of human interferon-a2b. Memari, Hamid Rajabi Ramanan, Ramakrishnan Nagasundara Ariff, Arbakariya The production of human interferon alpha2b (IFN-α2b) in two expression systems, tobacco (Nicotiana tabaccum) and Escherichia coli, was compared in various aspects such as safety, yield, quality of product and productivity. In the E. coli system, IFN-α2b was expressed under a pelB signal sequence and a T7lac promoter in a pET 26b(+) vector. The same gene was also cloned in expression plant vector (pCAMBIA1304) between cauliflower mosaic virus promoter (CaMV35S) and poly A termination region (Nos) and expressed in transgenic tobacco plants. The expression of protein in both systems was confirmed by western immunoblotting and the quantity of the protein was determined by immunoassay. The amount of periplasmic expression in E. coli was 60 µg/L of culture, while the amount of nuclear expression in the plant was 4.46 µg/kg of fresh leaves. The result of this study demonstrated that IFN-α2b was successfully expressed in periplasm of bacterial and plant systems. The limitations on the production of IFN-α2b by both systems are addressed and discussed to form the basis for the selection of the appropriate expression platform. SP Versita 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/13689/1/Comparison%20of%20expression%20systems%20for%20the%20production%20of%20human%20interferon.pdf Memari, Hamid Rajabi and Ramanan, Ramakrishnan Nagasundara and Ariff, Arbakariya (2010) Comparison of expression systems for the production of human interferon-a2b. Central European Journal of Biology, 5 (4). pp. 446-455. ISSN 1895-104X http://www.springer.com 10.2478/s11535-010-0036-y English
spellingShingle Memari, Hamid Rajabi
Ramanan, Ramakrishnan Nagasundara
Ariff, Arbakariya
Comparison of expression systems for the production of human interferon-a2b.
title Comparison of expression systems for the production of human interferon-a2b.
title_full Comparison of expression systems for the production of human interferon-a2b.
title_fullStr Comparison of expression systems for the production of human interferon-a2b.
title_full_unstemmed Comparison of expression systems for the production of human interferon-a2b.
title_short Comparison of expression systems for the production of human interferon-a2b.
title_sort comparison of expression systems for the production of human interferon-a2b.
url http://psasir.upm.edu.my/id/eprint/13689/
http://psasir.upm.edu.my/id/eprint/13689/
http://psasir.upm.edu.my/id/eprint/13689/
http://psasir.upm.edu.my/id/eprint/13689/1/Comparison%20of%20expression%20systems%20for%20the%20production%20of%20human%20interferon.pdf